Peptides derived from hmg-coa reductase and cosmetic and/or pharmaceutical composition containing same

ABSTRACT

The present invention relates to peptides derived from human HMG-CoA reductase of general formula (I): 
       R 1 -(AA) n -X1-Gly-Lys-X2-(AA) p -R 2 , 
     And of sequence SEQ ID No. 1 to SEQ ID No. 10. 
     The present invention also relates to a cosmetic or pharmaceutical composition comprising at least one peptide of general formula (I), in a physiologically suitable medium. 
     The present invention further relates to the use of this novel peptide as an active principle that activates human HMG-CoA reductase in a cosmetic composition intended to strengthen the barrier function of the skin and to stimulate epidermal differentiation. The invention further applies to a cosmetic treatment method intended to prevent and/or combat the external stresses and signs of cutaneous aging.

The present invention is situated in the cosmetic and pharmaceuticalfield, and more particularly in the dermatology field. The presentinvention relates to peptides derived from the human3-hydroxy-3-methyl-glutaryl Co-A reductase (human HMG-CoA reductase)enzyme of general formula (I):

R₁-(AA)_(n)-X1-Gly-Lys-X2-(AA)_(p)-R₂,

And of sequence SEQ ID No. 1 to SEQ ID No. 10.

The present invention also relates to a cosmetic or pharmaceuticalcomposition, comprising a peptide of general formula (I), used alone orin combination with at least one other active principle. The inventionis also relative to the use of this novel peptide as an active principlethat activates human HMG-CoA reductase in a cosmetic compositionintended to strengthen the barrier function of the skin and to stimulateepidermal differentiation. The invention also relates to the use of thisnovel active principle for producing a pharmaceutical composition, inparticular a dermatological composition, intended to prevent or treatpathological dysfunctions connected to an alteration in barrierfunction. The invention further applies to a cosmetic treatment processintended to prevent and/or combat the external stresses and cutaneoussigns of aging, according to which an effective quantity of activeprinciple, or a composition containing the active principle, is appliedto the areas to be treated.

The first function of the epidermis is to constitute a barrier betweenthe external environment and the internal environment. The outermostlayer of the epidermis, the horny layer of the epidermis, ensures thisfunction. It is composed of keratinocytes in the last stage of theirdifferentiation, corneocytes, sealed to each other by thickintercellular cement that is both flexible and impermeable. Therefore, acellular compartment constituted of corneocytes and an extracellularcompartment mainly constituted of lipids, organized into multilamellarstructures, are distinguished in the horny layer of the epidermis.

The lipid content of the human horny layer of the epidermis is estimatedat 15% cholesterol ester, 16% free long-chain saturated fatty acids, 32%cholesterol and 37% ceramides, although inter-individual variations arerather significant (Norlen L. et al. J. Invest. Dermatol. 1999; 112(1)p. 72-77). These lipids are synthesized by keratinocytes fromintermediate layers of the epidermis, and secreted in specializedorganites called “lamellar bodies” or Odland bodies. In particular, theepidermis is a very active site for synthesizing cholesterol. Therate-limiting step of this synthesis, and the most finely regulatedstep, is the conversion of the 3-hydroxy-3-methylglutaryl-Coenzyme A(HMG-CoA) into mevalonate. This step is catalyzed by a membrane-boundenzyme called HMG-CoA-reductase (E.C. 1.1.1.34). Human genome sequencingdata show that at least 2 isoforms for HMG-CoA reductase exist, coded bya unique gene, located on chromosome 5 (Luskey et al., J Biol. Chem.,1985 260(18), p. 10271-7).

In the skin, cholesterol plays a role in the fluidity of the membranesand, in particular, it seems to ensure the mobility of hydrocarbonchains in lipid bilayers (Martini M. C., Pathol. Biol. 2003, (51), p.267-270). Thus, in physiological situations, cholesterol is synthesizedat a level necessary for maintaining homeostasis. On the other hand,following a sudden alteration in the cutaneous barrier, a significantand rapid increase in the synthesis of cholesterol is observed,associated with an increase in the expression and activity of HMG-CoAreductase (Menon G. K. et al., J. Lipid, Res., 1985, (26), P. 418-427).

The key role of HMG-CoA reductase makes it a prime target for modulatingthe expression of cholesterol in the organism. Therefore, a class ofpharmacological compounds intended to inhibit HMG-CoA reductase has beendeveloped with the goal of lowering circulating cholesterol, calledstatins. This inhibitor effect of statins is also manifested in humanskin. In fact, the experimental administration of statins by topicalroute disrupts the barrier function of the skin (Proksch E. et al.,British J. Dermatol., 1993, (128), p. 473-482). These results confirmthe importance of cholesterol in the epidermal barrier function and thecentral role of HMG-CoA reductase in modulating its synthesis.

When skin ages, the integrity of the skin barrier as well as itscapacities for repair change. A global deficiency in lipids is observed,resulting in a reduction of lipid multilayers of the extracellularcompartment of the horny layer of the epidermis. These functionalchanges correlate with an increased susceptibility of aged skins toexternal stresses (Ghadially R. et al., J Clin Invest., 1995 (95 (5), p.2281-90).

Independently from intrinsic or photo-induced aging, alterations in theskin barrier may be produced during external stresses.

The expression “external stress” is understood to refer to stresses thatthe environment may produce. By way of example one may cite stressessuch as pollution, UV radiation or else irritating products such assurface active agents, preservatives or fragrances, or mechanicalstresses, such as abrasions, shaving or epilation. Pollution isunderstood to refer to both “external” pollution, due for example todiesel particles, ozone or heavy metals and to “internal” pollution,that may be particularly due to the emissions from paint, adhesive orwallpaper solvents (such as toluene, styrene, xylene or benzaldehyde),or else to cigarette smoke. Dryness of the atmosphere is also animportant cause of skin stress. These external stresses result in analteration of the barrier function that results in skin discomfort,disagreeable sensory phenomena, such as tearing pain or itching and evenexcessive fragility and redness.

In this context, trying to prevent the alteration or reestablish thebarrier function of the epidermis is desirable. In this particulardomain, the direct supply of lipid substitutes, such as ceramides (EP1272148, US2007576937) or certain cholesterol derivatives (FR 2 789 312)has largely been described. On the other hand, the utilization ofvegetable oils to activate the synthesis of cutaneous lipids has alsobeen described. (EP 1 707 189). In the cosmetics field, the moleculartargeting of HMG-CoA reductase has already been exploited in the goal ofinhibiting this key enzyme, for example by utilizing statins, alreadyknown for their HMGCOA-inhibiting properties, in the goal of obtainingan anti-aging effect (EP 0738510). However, to date, no documentdescribes or suggests that a peptide derived from human HMG-CoAreductase may have interesting properties to strengthen the barrierfunction of the skin and to stimulate epidermal differentiation.Activating HMG-CoA reductase in the goal of strengthening the barrierfunction and epidermal differentiation has now been contemplated. Bythis action, it is also possible to improve certain pathologicaldysfunctions connected to the barrier function (hypersensitive,irritated or reactive skin, atopic eczema).

The main objective of the present invention is to provide a novel activeprinciple, capable of strengthening the barrier function of the skin,stimulating epidermal differentiation and thus preventing signs of agingof the skin or protecting the skin from external stresses. In fact, theinventors have demonstrated a cosmetic and therapeutic, and particularlya dermatological activity of peptides derived from human HMG-CoAreductase.

In particular, it has been demonstrated that these peptides, whenapplied to the skin, strengthen the barrier function of the epidermisand stimulate epidermal differentiation. These properties have beendemonstrated by better protection of the skin tissue in relation toexternal stresses and by an increase in the production of lipidsconstituting the horny layer of the epidermis.

“Active principle that activates HMG-CoA reductase or is capable ofactivating human HMG-CoA” is understood to be any biologically activepeptide or derivative capable of increasing HMG-CoA reductase activity,either by increasing the protein synthesis of HMG-CoA reductase (bydirect or indirect modulation of the gene expression of HMG-CoAreductase), or by increasing the enzymatic activity of HMG-CoAreductase, or by other biological processes such as stabilization of theHMG-CoA reductase protein or else stabilization of messenger RNAtranscripts.

Skin is understood to refer to all of the covering tissues constitutingthe skin and mucous membranes.

“Topical application” is understood to refer to the act of applying orspreading the active principle according to the invention, or acomposition containing the principle, to or on the surface of the skin.

“Physiologically acceptable” is understood to mean that the activeprinciple according to the invention, or a composition containing theprinciple, is appropriate for entering in contact with the skin withoutcausing toxicity or intolerance reactions.

Thus, the first object of the invention is a peptide derived from humanHMG-CoA reductase.

The expression “peptide derived from human HMG-CoA reductase” designatesany biologically active peptide fragment in which the amino acidsequence is partially or entirely analogous or homologous to the humanHMG-CoA reductase peptide sequence.

The expression “biologically active” is understood to mean “has an invivo or in vitro activity characteristic of the activity of the activeprinciple according to the invention.”

According to a particularly advantageous embodiment of the invention,the peptide has a sequence that responds in part or in full to generalformula (I)

R₁-(AA)_(n)-X₁-Gly-Lys-X₂-(AA)_(p)-R₂

In which,

X₁ is lysine or arginine or any amino acid,

X₂ is serine or threonine,

AA represents any amino acid, or one of its derivatives, and n and p areintegers between 0 and 4,

R₁ represents the primary amine function of the N-terminal amino acid,free or substituted by a protecting group that may be chosen from amongan acetyl group, a benzoyl group, a tosyl group or a benzyloxycarbonylgroup,

R₂ represents the hydroxyl group of the carboxyl function of theC-terminal amino acid, free or substituted by a protecting group thatmay be chosen from among an alkyl chain from C₁ to C₂₀, or an NH2, NHYor NYY group with Y representing an alkyl chain from C₁ to C₄. Thepeptide is characterized in that the peptide is chosen from among thefollowing sequences:

(SEQ ID No. 1) Glu-Gly-Lys-Gly-Lys-Ser-Val-Val-Cys-Glu (SEQ ID No. 2)Glu-Gly-Lys-Gly-Lys-Ser-Val-Val (SEQ ID No. 3) Asp-Gly-Lys-Gly-Lys-Thr(SEQ ID No. 4) Arg-Gly-Lys-Ser (SEQ ID No. 5) Arg-Gly-Lys-Ser-NH₂(SEQ ID No. 6) Arg-Gly-Lys-Thr (SEQ ID No. 7) Arg-Gly-Lys-Thr-NH₂(SEQ ID No. 8) Lys-Gly-Lys-Ser (SEQ ID No. 9) Lys-Gly-Lys-Ser-NH₂(SEQ ID No. 10) Gly-Lys-Ser-NH₂

According to a particularly interesting embodiment, the biologicallyactive peptide corresponds to the SEQ ID No. 4 sequence.

According to another particularly interesting embodiment, thebiologically active peptide corresponds to the SEQ ID No. 5 sequence.

The invention also relates to homologous forms of these sequences. Theterm “homologous” designates, according to the invention, any peptidesequence identical to at least 50%, or preferably at least 80%, andstill more preferentially to at least 90% of said peptide sequence,chosen from among the SEQ ID No. 1 to SEQ ID No. 10 sequences. “Peptidesequence identical to at least X %” is understood to designate apercentage identity between the amino acid residues of two sequences tobe compared, obtained after the optimal alignment of the two sequences.The optimal alignment is obtained by using local homology algorithmssuch as those used by the BLAST P or T BLAST N computer softwareavailable on the NCBI site.

The term “homologous” may also designate a peptide that differs from thesequence of a peptide of SEQ ID No. 1 to SEQ ID No. 10 sequence by thesubstitution of chemically equivalent amino acids, i.e., by thesubstitution of a residue by another having the same characteristics.Thus, conventional substitutions take place between Ala, Val, Leu andIle; between Ser and Thr; between the acid residues Asp and Glu; betweenAsn and Gln; and between the basic residues Lys and Arg; Or between thearomatic residues Phe and Tyr.

In the invention, the term “amino acid” here refers to any natural ornon-natural organic acid having the formula:

—NHR—CR—C(O)—O—

Where each —R is independently selected between a hydrogen and an alkylgroup having between 1 and 12 carbon atoms. Preferentially, at least one—R group of each amino acid is a hydrogen. Here the term “alkyl” refersto a carbon chain that may be linear or branched, substituted (mono- orpoly-) or non-substituted; Saturated, monosaturated (a double or triplebond in the chain) or polyunsaturated (two or more double bonds, two ormore triple bonds, one or more double bonds and one or more triple bondsin the chain).

The term “peptide” designates a linkage of two or more amino acidsinterlinked by peptide linkages or by modified peptide linkages.

“Peptide” is also understood to refer to the natural or syntheticpeptide of the invention as described above, or at least one of itsfragments, whether obtained by proteolysis or synthetically, or else anynatural or synthetic peptide whose sequence is partially or totallyconstituted by the sequence of the peptide previously described.

So as to improve resistance to degradation, it may be necessary to use aprotected form of the peptide according to the invention. The form ofprotection must obviously be a biologically compatible form and must becompatible with a use in the field of cosmetics or pharmacy.

Many forms of biologically compatible protection may be contemplated.They are well known to the person skilled in the art as, for example,the acylation or acetylation of the amino terminal end, or the amidationor esterification of the carboxy terminal end. Thus, the inventionrelates to a composition such as previously defined, characterized bythe fact that the peptide of SEQ ID No. 1 to SEQ ID No. 10 is inprotected or unprotected form. Protection based on a substitution on theamino terminal end by an acetyl group, a benzoyl group, a tosyl group ora benzyloxycarbonyl group may be utilized. Preferably, protection basedon the amidation of the hydroxyl function of the carboxy terminal end byan NYY group with Y representing an alkyl chain from C₁ to C₄, or theesterification by an alkyl group is utilized. It is also possible toprotect the two ends of the peptide.

The peptide derivatives also relate to amino acids and peptidesinterconnected by a pseudo-peptidic linkage. “Pseudo-peptidic linkage”is understood to refer to all types of linkages capable of replacing“conventional” peptidic linkages.

In the domain of amino acids, the molecules have a geometry such thatthey may theoretically be present in the form of different opticalisomers. Thus, there exists a molecular conformation of the amino acid(AA) that rotates the plane of polarized light to the right(dextrorotatory conformation or D-aa), and a molecular conformation ofamino acid (aa) that rotates the plane of polarized light to the left(levorotatory conformation or L-aa). Natural amino acids are always oflevorotatory conformation; consequently, a peptide of natural originwill only be constituted of L-aa type amino acids. However, chemicalsynthesis in laboratory enables amino acids with the two possibleconformations to be prepared. From this base material, it is possible toincorporate, during peptide synthesis, amino acids in bothdextrorotatory and levorotatory optical isomer forms. Thus, amino acidsconstituting the peptide according to the invention may be in L- and D-configurations; preferentially, the amino acids are in L form. Thepeptide according to the invention may thus be in L-, D- or DL-form.

The peptide of general formula (I) according to the invention may beobtained either by conventional chemical synthesis (in solid phase or inhomogeneous liquid phase), or by enzymatic synthesis (Kullman et al., J.Biol. Chem. 1980, 225, 8234), from constituent amino acids or theirderivatives.

The peptide according to the invention may be of natural or syntheticorigin. Preferentially according to the invention, the peptide isobtained by chemical synthesis.

According to the invention, the active principle may be a singlepeptide, a mixture of peptides or peptide derivatives and/or constitutedof amino acid derivatives.

According to the invention, said peptide or mixture of peptides may beutilized as a medication.

According to an advantageous embodiment of the invention, the activeprinciple according to the invention is previously solubilized in one ormore physiologically acceptable solvents, conventionally used by theperson skilled in the art, such as water, glycerol, ethanol, propyleneglycol, butylene glycol, dipropylene glycol, ethoxylated diethyleneglycol or propoxylated diethylene glycol, cyclic polyols, whitepetroleum jelly, vegetable oil or any mixture of these solvents.

According to another advantageous embodiment of the invention, theactive principle according to the invention is previously solubilized ina cosmetic or pharmaceutical carrier such as liposomes, or adsorbed onpowdery organic polymers, mineral supports such as talcs and bentonites,and more generally solubilized in, or fixed on, any physiologicallyacceptable carrier.

The second object of the invention is a cosmetic, pharmaceutical, andparticularly dermatological composition, comprising a peptide of generalformula (I), as an active principle, used alone or in combination withat least one other active principle, in a physiologically suitablemedium.

Of course, it is obvious that the invention is aimed at mammals ingeneral, and more particularly at human beings.

According to an advantageous embodiment of the invention, the activeprinciple according to the invention is present in the compositions ofthe invention at a concentration of between approximately 0.0005 and 500ppm (parts per million), and preferentially at a concentration ofbetween approximately 0.01 and 5 ppm with relation to the total weightof the final composition.

The usable composition according to the invention may in particularconsist of a composition for hair care, and notably a shampoo, aconditioner, a treatment lotion, a styling cream or gel, a hairrestructuring lotion, a mask, etc. The cosmetic composition according tothe invention may be notably used in treatments implementing anapplication that is followed or not followed by rinsing or else inshampoo form. Thus, the active principle according to the invention mayadvantageously be utilized in antidandruff care of the scalp.

The active principle may also be present in the form of hair dye ormascara to be applied by brush or comb, in particular on the eyelashes,eyebrows or hair.

It is understood that the active principle according to the inventionmay be utilized alone or rather in combination with at least one otheractive principle, in a cosmetic composition or for the preparation of apharmaceutical and/or dermatological composition. Advantageously, theusable compositions according to the invention also contain variousprotective or anti-aging active principles intended, in particular, forthe prevention and/or treatment of disorders linked to aging.

The compositions according to the invention will be applied by anyappropriate route, notably oral, parenteral or external topical, andtheir formulations will be adapted by the person skilled in the art, inparticular for cosmetic or dermatological compositions. Advantageously,the compositions according to the invention are intended for topicaladministration on the skin. These compositions therefore must contain aphysiologically acceptable medium, i.e., a medium compatible with theskin and epithelial appendages, and must cover all cosmetic ordermatological forms. These compositions will notably be in the form ofcreams, oil in water emulsions, or water in oil emulsions or multipleemulsions, solutions, suspensions, gels, milks, lotions, sticks or elsepowders, and suitable for an application on the skin, lips and/orepithelial appendages. These compositions comprise the excipientsnecessary for their formulation, such as solvents, thickeners, diluents,surface active agents, antioxidants, colorants, preservatives andfragrances.

According to another embodiment of the invention, the compositions willbe appropriate for oral administration for pharmaceutical use. Thus, thecompositions may in particular be present in the form of tablets,capsules, gel capsules, chewable pastes, powders to consume as is or tobe mixed immediately before use with a liquid, syrups, gels or any otherform known to the person skilled in the art. They will contain suitableformulation excipients, such as colorants, sweeteners, flavorings,bulking agents, binders and preservatives.

These compositions may particularly be present in the form of an aqueoussolution, hydroalcoholic or oily solution; an oil in water emulsion,water in oil emulsion or multiple emulsions; They may also be present inthe form of creams, suspensions or else powders, suitable forapplication on the skin, mucous membranes, lips and/or epithelialappendages. These compositions may be more or less fluid and have theappearance of a cream, lotion, milk, serum, pomade, gel, paste or foam.They may also be present in solid form, such as a stick, or may beapplied on the skin in aerosol form. They may be utilized as a careproduct and/or as a skin makeup product.

These compositions also comprise any additive commonly utilized in thecontemplated field of application as well as the adjuvants necessary fortheir formulation, such as solvents, thickeners, diluents, antioxidants,colorants, sunscreens, self-tanning agents, pigments, fillers,preservatives, fragrances, odor absorbers, cosmetic or pharmaceuticalactive ingredients, essential oils, vitamins, essential fatty acids,surface active agents, film-forming polymers, etc.

In all cases, the person skilled in the art will make sure that theseadjuvants as well as their proportions are chosen so as to not harm thedesired advantageous properties of the composition according to theinvention. These adjuvants may, for example, correspond to 0.01 to 20%of the total weight of the composition. When the composition of theinvention is an emulsion, the fatty phase may represent from 5 to 80% byweight and preferably from 5 to 50% by weight with relation to the totalweight of the composition. The emulsifiers and co-emulsifiers utilizedin the composition will be chosen from among those conventionallyutilized in the field under consideration. For example, they may beutilized in a proportion going from 0.3 to 30% by weight with relationto the total weight of the composition.

The third object of the invention is the utilization, in a cosmeticcomposition, of an effective quantity of peptide of general formula (I),as an active principle that activates human HMG-CoA reductase.

The effective quantity of active principle corresponds to the quantitynecessary for obtaining the desired result, that is, to activate theHMG-CoA reductase, in the goal of improving the barrier function of theepidermis and stimulating epidermal differentiation.

“Strengthen the barrier function of the skin and stimulate epidermaldifferentiation” is understood to refer to the improvement in theprotection capacity of the horny layer of the epidermis and the increasein the expression of biological differentiation markers, such askeratins.

Thus, thanks to the special properties of said active principle, it canbe used in a cosmetic composition intended to strengthen the barrierfunction of the skin and to stimulate epidermal differentiation.

On the other hand, the peptide of general formula (I) can be usedadvantageously as an active principle in a cosmetic composition intendedto preventively and/or curatively combat the signs of cutaneous aging,and more particularly photo-induced aging (photo aging). Cutaneous signsof aging is understood to refer to any modifications in the externalappearance of the skin and epithelial appendages due to aging such as,for example, superficial roughness of the horny layer of the epidermis,wrinkles and fine lines, but also any internal modification of the skinthat is not systematically manifested in a modified external appearancesuch as, for example, thinning of the dermis or any other internaldegradation of the skin following exposure to ultraviolet (UV)radiation.

According to another aspect of the invention, the peptide of generalformula (I) can be used advantageously as an active principle in acosmetic composition intended to protect the skin against all types ofexternal stresses.

In particular, the object of the invention is the utilization of acosmetic composition comprising an effective quantity of peptideaccording to the invention to prevent or treat damage caused to the skinby mechanical treatments such as shaving or epilation.

In particular, the object of the invention is the utilization of acosmetic composition comprising an effective quantity of peptideaccording to the invention to prevent or treat damage caused to the skinby extreme climactic conditions or sudden variations in temperatures andhygrometry.

The invention further consists of the utilization of the peptideaccording to the invention for preparing a pharmaceutical compositionintended to prevent or treat pathologies characterized by an alterationin the barrier function, such as hypersensitive, irritated or reactiveskin and atopic eczema.

The invention further consists of a cosmetic treatment method intendedto prevent and/or combat external stresses according to which acomposition comprising an effective quantity of peptide according to theinvention is applied onto the areas to be treated.

The invention further consists of a cosmetic treatment method intendedto prevent and/or combat cutaneous signs of aging and/or photo-aging,according to which a composition comprising an effective quantity ofpeptide according to the invention is applied onto the areas to betreated.

Particular embodiments of this cosmetic treatment method also resultfrom the previous description. Other advantages and characteristics ofthe invention will more clearly appear upon reading the examples givenfor illustrative and non-limiting purposes.

EXAMPLE 1 Ultrastructural Study of Lamellar Bodies in HumanKeratinocytes Treated by Peptide SEQ ID No. 5

The goal of this study is to study in an ultrastructural manner, intransmission electron microscopy, keratinocytes treated by peptide SEQID No. 5 at 0.5 ppm.

Protocol:

Normal human keratinocytes in culture are treated with a 1% stocksolution at 50 ppm of peptide SEQ ID No. 5 for 48 hours (the medium inthe presence of the active ingredient is changed every 24 hours). Thecells are washed in PBS, and then are fixed by Karnosky hypertonicfixation (4% paraformaldehyde, 5% glutaraldehyde in a 0.08M phosphatebuffer) 1 hour at ambient temperature and then 24 hours at 4° C. Thecells are detached from the support by scraping and centrifuged 5minutes at 1000 rpm. The supernatant is eliminated and a 1M sodiumcacodylate buffer is deposited on the residue. The cells are mixed with2% agar and then postfixed by osmium tetraoxide for 1 hour. Thespecimens are then dehydrated by successive passages in a series ofalcohol (from 50 to 100%). The cells are then coated in a resin. Thepolymerization is carried out for approximately 12 hours at 60° C.Semi-thin sections of 0.5 μm are made with an ultramicrotome. Thesections are deposited on a heat bonded slide and then colored withtoluidine blue. The slides are then dehydrated again and mounted in asuitable medium. After having chosen the optimal study zone, the blockis recut to the desired size and ultrathin sections are then made, onlysections with a silver-grey color and a suitable size are mounted on theelectron microscopy grid labeled with both uranyl acetate and leadcitrate, and are examined by transmission electron microscope at 60 or80 KV.

Results: The ultrastructural study shows that the Golgi complex issubstantially more developed than in the control cells. This increase isconnected to an excess production of lamellar bodies (or Odland bodies)that is the sign of an increase in lipid synthesis.

Conclusions: Peptide SEQ ID No. 5 at 0.5 ppm is capable of inducing anincrease in lipid synthesis in normal human keratinocytes.

EXAMPLE 2 Ultrastructural Study of Caveolae in Human Fibroblasts Treatedby Peptide SEQ ID No. 4

The goal of this study is to study at the ultrastructural level thecaveolae in human dermal fibroblasts.

Protocol: Normal human dermal fibroblasts in culture are treated with a1% stock solution at 50 ppm of peptide SEQ ID No. 4 for 48 hours (themedium in the presence of the active ingredient is changed every 24hours).

Results: The ultrastructural study shows a significant increase incaveolae in cells treated by peptide SEQ ID No. 4, in comparison withuntreated control cells. These results are the sign of a positive effectof the active principle, since the caveolae are invaginations of theplasma membrane that enable the externalization of molecules such ascholesterol.

Conclusions: Peptide SEQ ID No. 4 at 0.5 ppm causes the increase inmembrane structures with externalization of cholesterol.

EXAMPLE 3 Differentiation Study of Human Keratinocytes Treated byPeptide SEQ ID No. 5

The goal of this study is to determine the influence of peptide SEQ IDNo. 5 on epidermal differentiation.

Protocol: Normal human keratinocytes in culture are treated with a 1%stock solution at 50 ppm of peptide SEQ ID No. 4 for 48 hours (themedium in the presence of the active ingredient is changed every 24hours). The cells are then washed and fixed with cold methanol for 4minutes at 4° C. The cells are incubated in the presence of a monoclonalanti-cytopankeratin antibody at 1:200 for 1 hour at ambient temperatureand are then revealed by a second antibody at 1:50 for 1 hour at ambienttemperature, coupled with a fluorescent dye, “alexa 488.” After mountingin a particular medium, the slides are observed by epifluorescencemicroscope.

Results: The active ingredient increases the expression of pankeratinsin the treated cells.

Conclusions: Peptide SEQ ID No. 5, at 0.5 ppm, increases the expressionof pankeratins in normal human keratinocytes. In the presence of peptideSEQ ID No. 5, the cells are stimulated in pathway differentiation.

EXAMPLE 4 Study of the Protective Effect of Peptide SEQ ID No. 5 on SkinCells Subjected to Ultraviolet Radiation (UVB)

The goal of this study is to determine the protective effect of peptideSEQ ID No. 5 with relation to normal human keratinocytes subjected tostress by UVB radiation. To do this, cellular viability tests wereconducted by the MTT technique.

Protocol: The normal human keratinocytes are treated with a 1% solution,a solution at 50 ppm of peptide SEQ ID No. 5, for 24 hours, irradiatedby UVB (50 mJ/cm²) and then cultivated again 24 hours in the presence ofthe same concentration of peptide SEQ ID No. 5. Untreated and irradiatedcontrols are carried out under the same conditions. At the end of theexperiment, the cells are incubated in a solution containing 0.1 mg/mlof MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide).This compound is absorbed by the living cells and then metabolized bymitochondrial enzymes into a blue violet compound, formazan, that willbe assayed by spectrophotometry at 540 nm. The optical density (O.D.) Isthen directly proportional to the mitochondrial enzymatic activity aswell as to the number of living cells.

Results: Evaluation of cellular viability by the MTT technique showsthat peptide SEQ ID No. 5 increases cellular viability after UVBirradiation by 16%.

Conclusions: Peptide SEQ ID No. 5, at the 0.5 ppm concentration,increases cellular viability and effectively protects the skin cellsfrom the cytotoxic effects of UVB radiation.

EXAMPLE 5 Study of the Expression of HMG-CoA Reductase in Skin Biopsiesin the Presence of Peptide SEQ ID No. 5

The goal of this study is to determine the influence of peptide SEQ IDNo. 5 on the expression of HMG-CoA reductase.

Protocol: Samples of human skin are placed in culture at the air/liquidinterface. A 1% stock solution at 50 ppm of peptide SEQ ID No. 5 isapplied topically and then the samples are incubated for 24 hours or 48hours.

These skin samples are then fixed with formaldehyde and then enclosed inparaffin. Sections of 2 to 3 μm are then made. Immunolabelling iscarried out after unmasking the specific sites by microwave treatmentand then incubation in trypsin. Immunolabelling is carried out by usinga polyclonal rabbit antibody specific for HMG-CoA reductase (Millipore,Upstate), and then a secondary antibody, coupled with a fluorescent dye.The skin sections are then examined by epifluorescence microscope (NikonEclipse E600 microscope).

Results: Microscopic observations show stronger fluorescence in skintreated by peptide SEQ ID No. 5, in the upper layers of the epidermis,with relation to the untreated control.

Conclusions: Peptide SEQ ID No. 5, at the 0.5 ppm concentration,stimulates the expression of HMG-CoA reductase, in the upper layers ofthe epidermis.

EXAMPLE 6 Study of the Expression of HMG-CoA Reductase in Normal HumanKeratinocytes in the Presence of Peptide SEQ ID No. 4

The goal of this study is to determine the influence of peptide SEQ IDNo. 4 on the expression of HMG-CoA reductase in normal humankeratinocytes.

Protocol: Normal human dermal keratinocytes in culture are treated witha 1% stock solution at 50 ppm of peptide SEQ ID No. 4 for 48 hours (themedium in the presence of the active ingredient is changed every 24hours). The cells are then washed and fixed in cold methanol for 4minutes at 4° C. The cells are incubated in the presence of a polyclonalrabbit antibody specific for HMG-CoA reductase (Millipore, Upstate), andthen a secondary antibody coupled with a fluorescent dye. The cells arethen examined by epifluorescence microscope (Nikon Eclipse E600microscope).

Results: Microscopic observations show more intense cytoplasmicfluorescence in cells treated by peptide SEQ ID No. 4.

Conclusions: Peptide SEQ ID No. 4, at the 0.5 ppm concentration,stimulates the expression of HMG-CoA reductase in normal humankeratinocytes.

EXAMPLE 7 Preparation of Compositions

1-Sun Protection Cream:

Weight Trade names INCI names percent PHASE A Demineralized water Aqua(Water) qsp Pemulen TR1 Acrylates/C10-30 Alkyl 0.40 AcrylateCrosspolymer Glycerine Glycerin 3.00 Nipastat Sodium SodiumMethylparaben (and) 0.15 Sodium Ethylparaben (and) Sodium Butyl paraben(and) Sodium Propylparaben (and) Sodium Isobutylparaben PHASE B ParsolMCX Ethylhexyl Methoxycinnamate 7.50 Eusolex 4360 Benzophenone-3 3.00Parsol 1789 Butyl Methoxydibenzoyl- 2.00 methane Myritol 318Caprylic/Capric Triglyceride 4.00 Emulgade SEV Hydrogenated PalmGlycerides 5.00 (and) Ceteareth-20 (and) Ceteareth-12 (and) CetearylAlcohol Propylparaben Propylparaben 0.15 Nacol 16-98 Cetyl Alcohol 1.00PHASE C TEA Triethanolamine 0.20 PHASE D Peptide SEQ ID No. 4 3 ppmFragrance Fragrance qsp Colorant qsp

The constituents of phase A and phase B are heated separately between70° C. and 75° C. Phase B is emulsified in phase A under stirring. PhaseC is added at 45° C., by increasing the stirring. Phase D is then addedwhen the temperature is below 40° C. The cooling is continued until 25°C. under intensive stirring.

2-Anti-Age Cream:

Weight Trade names INCI names percent Phase A Montanov 68 CetearylAlcohol (and) 6.00 Cetearyl Glucoside Squalane Squalane 3.00 Cetiol SB45 Butyrospermum Parkii (Shea 2.00 Butter) Waglinol 250 CetearylEthylhexanoate 3.00 Amerchol L- 101 Mineral Oil (and) Lanolin 2.00Alcohol Abil 350 Dimethicone 1.50 BHT BHT 0.01 Coenzyme Q10 Ubiquinone0.10 Phase B Avocado oil Persea Gratissima (Avocado) 1.25 Oil PhenonipPhenoxyethanol (and) 0.75 Methylparaben (and) Ethylparaben (and)Butylparaben (and) Propylparaben (and) Isobutylparaben Phase CDemineralized water Aqua (Water) qsp Butylene Glycol Butylene Glycol2.00 Glucam E10 Methyl Gluceth-10 1.00 Allantoin Allantoin 0.15 CarbopolUltrez 10 Carbomer 0.20 Phase D TEA Triethanolamine 0.18 Phase E PeptideSEQ ID No. 4 0.5 ppm GP4G Water (and) Artemia Extract 1.50 CollaxylWater (and) Butylene Glycol 3.00 (and) Hexapeptide-9 Phase F FragranceFragrance qsp Colorant qsp

Prepare and melt phase A at 65-70° C. Heat phase C to 65-70° C. Phase Bis added to phase A just before emulsifying A into B. At approximately45° C., the carbomer is neutralized by adding phase D. Phase E is thenadded under mild stirring and cooling is continued until 25° C. Phase Fis then added if desired.

3-Protective Day Cream:

Weight Trade names INCI names percent Phase A Emulium Delta Cetylalcohol (and) Glyceryl 4.00 Stearate (and) PEG-75 Stearate (and)Ceteth-20 (and) Steareth-20 Lanette O Cetearyl Alcohol 1.50 D C 200Fluid/100 cs Dimethicone 1.00 DUB 810C Coco Caprylate/Caprate 1.00 DPPGPropylene Glycol 3.00 Dipelargonate DUB DPHCC Dipentaerythrityl 1.50Hexacaprylate/Hexacaprate Cegesoft PS6 Vegetable Oil 1.00 Vitamin ETocopherol 0.30 Phenonip Phenoxyethanol (and) 0.70 Methylparaben (and)Ethylparaben (and) Butylparaben (and) Propylparaben (and)Isobutylparaben Phase B Demineralized water Aqua qsp 100 GlycerineGlycerin 2.00 Carbopol ETD 2020 Acrylates/C10-30Alkyl 0.15 AcrylateCrosspolymer Keltrol BT Xanthan Gum 0.30 Phase C Sodium Hydroxide SodiumHydroxide 0.30 (sol. 10%) Phase D Demineralized water Aqua 5.00 Stay-C50 Sodium Ascorbyl Phosphate 0.50 Phase E Butylene Glycol ButyleneGlycol 2.00 Dekaben CP Chlorphenesin 0.20 Phase F GP4G Water (and)Artemia Extract 1.00 Peptide SEQ ID No. 5 5 ppm

Prepare phase A and heat to 75° C. under stirring. Prepare phase B bydispersing the carbopol and then the xanthan gum under stirring. Letrest. Heat to 75° C.

At temperature, emulsify A into B under rotor stator stirring.Neutralize with phase C under rapid stirring. After cooling to 40° C.,add phase D, and then phase E. Cooling is continued under mild stirringand phase F is added.

1. A peptide derived from human HMG-CoA reductase, of general formula(I)R₁-(AA)_(n)-X₁-Gly-Lys-X₂-(AA)_(p)-R₂ in which, X₁ is lysine or arginineor any amino acid, X₂ is serine or threonine, AA represents any aminoacid, or one of its derivatives, and n and p are integers between 0 and4, R₁ represents the primary amine function of the N-terminal aminoacid, free or substituted by a protecting group that may be chosen fromamong an acetyl group, a benzoyl group, a tosyl group or abenzyloxycarbonyl group, R₂ represents the hydroxyl group of thecarboxyl function of the C-terminal amino acid, free or substituted by aprotecting group that may be chosen from among an alkyl chain from C₁ toC₂₀, or an NH₂, NHY or NYY group with Y representing an alkyl chain fromC₁ to C₄. characterized in that the peptide is chosen from among thefollowing sequences: (SEQ ID No. 1)Glu-Gly-Lys-Gly-Lys-Ser-Val-Val-Cys-Glu (SEQ ID No. 2)Glu-Gly-Lys-Gly-Lys-Ser-Val-Val (SEQ ID No. 3) Asp-Gly-Lys-Gly-Lys-Thr(SEQ ID No. 4) Arg-Gly-Lys-Ser (SEQ ID No. 5) Arg-Gly-Lys-Ser-NH₂(SEQ ID No. 6) Arg-Gly-Lys-Thr (SEQ ID No. 7) Arg-Gly-Lys-Thr-NH₂(SEQ ID No. 8) Lys-Gly-Lys-Ser (SEQ ID No. 9) Lys-Gly-Lys-Ser-NH₂(SEQ ID No. 10) Gly-Lys-Ser-NH₂.


2. The peptide according to claim 1 characterized in that the peptidecorresponds to the SEQ ID No. 4 sequence.
 3. The peptide according toclaim 1 characterized in that the peptide corresponds to the SEQ ID No.5 sequence.
 4. The peptide according to claim 1, wherein the peptide issolubilized in one or more physiologically acceptable solvents selectedfrom the group consisting of water, glycerol, ethanol, propylene glycol,butylene glycol, dipropylene glycol, ethoxylated diethylene glycol orpropoxylated diethylene glycol, cyclic polyols, white petroleum jelly,vegetable oil and combinations thereof.
 5. The peptide according toclaim 1, wherein the peptide is characterized as a medication.
 6. Acosmetic or pharmaceutical composition characterized in that thecomposition comprises, at least one peptide derived from human HMG-CoAreductase, of general formula (I)R₁-(AA)_(n)-X₁-Gly-Lys-X₂-(AA)_(p)-R₂ in which, X₁ is lysine or arginineor any amino acid, X₂ is serine or threonine, AA represents any aminoacid, or one of its derivatives, and n and p are integers between 0 and4, R₁ represents the primary amine function of the N-terminal aminoacid, free or substituted by a protecting group that may be chosen fromamong an acetyl group, a benzoyl group, a tosyl group or abenzyloxycarbonyl group, R₂ represents the hydroxyl group of thecarboxyl function of the C-terminal amino acid, free or substituted by aprotecting group that may be chosen from among an alkyl chain from C₁ toC₂₀, or an NH₂, NHY or NYY group with Y representing an alkyl chain fromC₁ to C₄. characterized in that the peptide is chosen from among thefollowing sequences: (SEQ ID No. 1)Glu-Gly-Lys-Gly-Lys-Ser-Val-Val-Cys-Glu (SEQ ID No. 2)Glu-Gly-Lys-Gly-Lys-Ser-Val-Val (SEQ ID No. 3) Asp-Gly-Lys-Gly-Lys-Thr(SEQ ID No. 4) Arg-Gly-Lys-Ser (SEQ ID No. 5) Arg-Gly-Lys-Ser-NH₂ (SEQID No. 6) Arg-Gly-Lys-Thr (SEQ ID No. 7) Arg-Gly-Lys-Thr-NH₂ (SEQ ID No.8) Lys-Gly-Lys-Ser (SEQ ID No. 9) Lys-Gly-Lys-Ser-NH₂ (SEQ ID No. 10)Gly-Lys-Ser-NH₂; and a physiologically acceptable medium; wherein thepeptide is present in the medium, as an active, alone or in combinationwith at least one other active principle.
 7. The cosmetic orpharmaceutical composition according to claim 6, characterized in thatsaid peptide is present at a concentration of between 0.0005 and 500ppm.
 8. The cosmetic or pharmaceutical composition according to claim 7,characterized in that said peptide is present at a concentration ofbetween 0.01 and 5 ppm.
 9. The cosmetic or pharmaceutical compositionaccording to claim 6 characterized in that the composition is atopically administrable composition.
 10. The cosmetic compositionaccording to claim 6, comprising an effective quantity of the peptide asan active that activates human HMG-CoA reductase.
 11. The cosmeticcomposition according to claim 10, wherein the cosmetic compositionstrengthens the barrier function of the epidermis and stimulatesepidermal differentiation.
 12. The cosmetic composition according toclaim 10, wherein the cosmetic composition prevents and combats thecutaneous signs of aging and photo aging.
 13. The cosmetic compositionaccording to claim 10, wherein the cosmetic composition protects theskin from external stresses.
 14. The cosmetic composition according toclaim 6, wherein the composition prevents or treats pathologiescharacterized by an alteration in the barrier function, such ashypersensitive, irritated or reactive skin and atopic eczema.
 15. Acosmetic treatment method intended to prevent and/or combat externalaggressions or cutaneous signs of aging and photo aging, the methodcomprising: topically applying, to skin to be treated, a compositioncomprising an effective quantity of a peptide derived from human HMG-CoAreductase, of general formula (I)R₁-(AA)_(n)-X₁-Gly-Lys-X₂-(AA)_(p)-R₂ in which, X₁ is lysine or arginineor any amino acid, X₂ is serine or threonine, AA represents any aminoacid, or one of its derivatives, and n and p are integers between 0 and4, R₁ represents the primary amine function of the N-terminal aminoacid, free or substituted by a protecting group that may be chosen fromamong an acetyl group, a benzoyl group, a tosyl group or abenzyloxycarbonyl group, R₂ represents the hydroxyl group of thecarboxyl function of the C-terminal amino acid, free or substituted by aprotecting group that may be chosen from among an alkyl chain from C₁ toC₂₀, or an NH₂, NHY or NYY group with Y representing an alkyl chain fromC₁ to C₄. characterized in that the peptide is chosen from among thefollowing sequences: (SEQ ID No. 1)Glu-Gly-Lys-Gly-Lys-Ser-Val-Val-Cys-Glu (SEQ ID No. 2)Glu-Gly-Lys-Gly-Lys-Ser-Val-Val (SEQ ID No. 3) Asp-Gly-Lys-Gly-Lys-Thr(SEQ ID No. 4) Arg-Gly-Lys-Ser (SEQ ID No. 5) Arg-Gly-Lys-Ser-NH₂(SEQ ID No. 6) Arg-Gly-Lys-Thr (SEQ ID No. 7) Arg-Gly-Lys-Thr-NH₂(SEQ ID No. 8) Lys-Gly-Lys-Ser (SEQ ID No. 9) Lys-Gly-Lys-Ser-NH₂(SEQ ID No. 10) Gly-Lys-Ser-NH₂ .


16. (canceled)
 17. The cosmetic treatment method according to claim 15,wherein the peptide is solubilized in one or more physiologicallysuitable solvents selected from the group consisting of water, glycerol,ethanol, propanediol, propylene glycol, butylene glycol, dipropyleneglycol, ethoxylated diethylene glycol or propoxylated diethylene glycol,cyclic polyols, white petroleum jelly, vegetable oil, and combinationsthereof.
 18. The cosmetic treatment method according to claim 15,wherein the peptide is present at a concentration of between 0.0005 and500 ppm.
 19. The cosmetic treatment method according to claim 15,wherein the peptide is present at a concentration of between 0.01 and 5ppm.